A direct fluorometric assay of benzo[a]pyrene hydroxylase. |
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| Authors: | C S Yang L P Kicha |
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| Affiliation: | Institut für Biochemie, Gesellschaft für Strahlen- und Umweltforschung, 8 München 2, Landwehrstr. 61, Germany |
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| Abstract: | A technique to measure the activity of pyruvate carboxylase spectrophotometrically in crude liver homogenates is described. The assay is based on the transformation of oxaloacetate, which is formed during the carboxylation reaction, into citrate in the presence of excess acetyl CoA and citrate synthase. After removal of pyruvate with KBH4 and of protein with HClO4, citrate is cleaved with citrate lyase into oxaloacetate and acetate, and oxaloacetate then is measured spectrophotometrically. Optimal concentrations of pyruvate, Mg2+, ATP, and KHCO3 for the carboxylation reaction and the Vmax were in good correlation with the data found by others using [14C]pyruvate. |
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| Keywords: | To whom correspondence should be addressed. |
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